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Molecular and Cellular Biology, May 2001, p. 3144-3158, Vol. 21, No. 9
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.9.3144-3158.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Saccharomyces cerevisiae CTF18 and CTF4 Are Required for Sister Chromatid Cohesion

Joseph S. Hanna,1 Evgueny S. Kroll,2 Victoria Lundblad,3 and Forrest A. Spencer1,*

McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland1; Molecular Sciences Institute, Berkeley, California2; and Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas3

Received 4 December 2000/Returned for modification 17 January 2001/Accepted 8 February 2001

CTF4 and CTF18 are required for high-fidelity chromosome segregation. Both exhibit genetic and physical ties to replication fork constituents. We find that absence of either CTF4 or CTF18 causes sister chromatid cohesion failure and leads to a preanaphase accumulation of cells that depends on the spindle assembly checkpoint. The physical and genetic interactions between CTF4, CTF18, and core components of replication fork complexes observed in this study and others suggest that both gene products act in association with the replication fork to facilitate sister chromatid cohesion. We find that Ctf18p, an RFC1-like protein, directly interacts with Rfc2p, Rfc3p, Rfc4p, and Rfc5p. However, Ctf18p is not a component of biochemically purified proliferating cell nuclear antigen loading RF-C, suggesting the presence of a discrete complex containing Ctf18p, Rfc2p, Rfc3p, Rfc4p, and Rfc5p. Recent identification and characterization of the budding yeast polymerase kappa , encoded by TRF4, strongly supports a hypothesis that the DNA replication machinery is required for proper sister chromatid cohesion. Analogous to the polymerase switching role of the bacterial and human RF-C complexes, we propose that budding yeast RF-CCTF18 may be involved in a polymerase switch event that facilities sister chromatid cohesion. The requirement for CTF4 and CTF18 in robust cohesion identifies novel roles for replication accessory proteins in this process.

* Corresponding author. Mailing address: Johns Hopkins University School of Medicine, 720 Rutland Ave./Ross 850, Baltimore, MD 21205. Phone: (410) 614-2536. Fax: (410) 955-0484. E-mail: fspencer{at}jhmi.edu.

Molecular and Cellular Biology, May 2001, p. 3144-3158, Vol. 21, No. 9
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.9.3144-3158.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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